内皮抑素和平阳霉素对血管内皮细胞的作用

发布时间：2018-06-17 01:54

本文选题：重组人血管内皮抑制素 + 平阳霉素　；参考：《广西医科大学》2011年硕士论文

【摘要】：目的：比较重组人血管内皮抑制素、平阳霉素单独使用及联合应用对人脐静脉内皮细胞系HUVEC细胞的作用效果,探讨相关作用机制,以揭示内皮抑素应用于增生期血管瘤治疗的可行性。方法：传代培养HUVEC细胞,取3-5代生长良好的细胞,将不同药物浓度内皮抑素、平阳霉素分别作用于HUVEC细胞,以四唑盐比色法(MTT法)比较两药作用24h、48h后对HUVEC细胞的细胞抑制率,苏木精-伊红(HE)染色光镜下观察药物作用前后细胞形态变化,用免疫组化法检测药物作用前后HUVEC细胞内血管内皮生长因子(VEGF)阳性表达的差异,流式细胞仪分析药物作用后HUVEC细胞的细胞凋亡情况。结果：内皮抑素、平阳霉素分别作用HUVEC细胞后的吸光度值(OD值)均随药物浓度增大而减小,两药对HUVEC细胞的抑制作用表现出一定的浓度依赖性和时间依赖性。VEGF免疫组化染色显示两种药物分别作用后HUVEC细胞的细胞胞浆灰度值均增大。光镜下可见：内皮抑素作用后HUVEC细胞形态明显改变,盖玻片上可看到大量凋亡细胞,并随药物浓度增大而增多,平阳霉素作用后大量细胞悬浮,贴壁细胞数量明显减少,药物浓度越大而贴壁细胞越少。流式细胞仪测得两种药物单独使用均会引起HUVEC细胞凋亡,且均表现为浓度越大凋亡细胞数越多；两种药物联合使用表现出对HUVEC细胞的协同抑制效应。结论：内皮抑素、平阳霉素对HUVEC细胞均有明显抑制作用,均能引起细胞凋亡,均阻止VEGF进入HUVEC细胞内从而抑制细胞的增殖,说明两药对血管内皮细胞的抑制作用的相关机制存在某种共性；较高浓度的内皮抑素仍以细胞凋亡为主,而平阳霉素浓度大于一定范围(20μg/ml)细胞抑制逐渐以坏死占主导；平阳霉素对血管内皮细胞贴壁的影响较内皮抑素明显,揭示两种药物对血管内皮细胞的抑制作用存在差异；内皮抑素与平阳霉素的联合使用对内皮细胞的抑制有协同作用,为两种药物联合使用治疗增殖期血管瘤提供基础理论依据。
[Abstract]:Aim: to compare the effects of recombinant human vascular endotheliostatin and pingyangmycin on human umbilical vein endothelial cell line HUVEC. To explore the feasibility of endostatin in the treatment of hyperplastic hemangioma. Methods: HUVEC cells were cultured and cultured in 3-5 passages. Different concentrations of endostatin and pingyangmycin were used to treat HUVEC cells respectively. The inhibitory rates of the two drugs on HUVEC cells after 24 h or 48 h treatment were compared by MTT assay. The morphological changes of HUVEC cells were observed under light microscope with hematoxylin and eosin (HEH) staining. The positive expression of vascular endothelial growth factor (VEGF) in HUVEC cells was detected by immunohistochemical method. Apoptosis of HUVEC cells was analyzed by flow cytometry. Results: the absorbance value and OD value of HUVEC cells treated with endostatin and pingyangmycin decreased with the increase of drug concentration. The inhibitory effects of the two drugs on HUVEC cells were concentration-dependent and time-dependent. VEGF immunohistochemical staining showed that the cytoplasmic gray values of HUVEC cells increased after the two drugs were treated separately. Under the light microscope, the morphology of HUVEC cells was obviously changed after the treatment of endostatin, a large number of apoptotic cells could be seen on the cover glass and increased with the increase of drug concentration. After the treatment of pingyangmycin, a large number of cells suspended and the number of adherent cells decreased obviously. The higher the drug concentration, the fewer adherent cells. The results of flow cytometry showed that both drugs alone could induce apoptosis of HUVEC cells, and the higher the concentration of the two drugs, the more the number of apoptotic cells, and the synergistic inhibitory effect of the two drugs on HUVEC cells. Conclusion: both endostatin and pingyangmycin can inhibit the proliferation of HUVEC cells, induce apoptosis, prevent VEGF from entering into HUVEC cells and inhibit the proliferation of HUVEC cells. The results showed that there was some common mechanism in the inhibition of vascular endothelial cells by the two drugs, the higher concentration of endostatin was still mainly apoptosis, while the inhibition of pingyangmycin was more than 20 渭 g / ml. The effect of pingyangmycin on the adhesion of vascular endothelial cells was more obvious than that on endostatin, which revealed that the inhibitory effects of two drugs on vascular endothelial cells were different, and the combination of endostatin and pingyangmycin had synergistic effects on the inhibition of endothelial cells. To provide basic theoretical basis for the combined use of two drugs in the treatment of proliferative hemangioma.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329.3

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